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King Saud University College of Science Department of Biochemistry Disclaimer The texts, tables and images contained in this course presentation are not my own, they can be found on: References supplied Atlases or The web Cellular Biochemistry BCH 102 Professor A. S. Alhomida Cellular Biochemistry: Cell Structure & Function THE CELL How We Study Cells 1. Microscopes provide windows to the world of the cell 2. Cell biologists can isolate organelles to study their function Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Microscopes provide windows to the world of the cell The discovery and early study of cells progressed with the invention and improvement of microscopes in the 17th century. In a light microscope (LMs) visible light passes through the specimen and then through glass lenses. The lenses refract light such that the image is magnified into the eye or a video screen. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Microscopes vary in magnification and resolving power. Magnification is the ratio of an object’s image to its real size. Resolving power is a measure of image clarity. It is the minimum distance two points can be separated and still viewed as two separate points. Resolution is limited by the shortest wavelength of the source, in this case light. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings The minimum resolution of a light microscope is about 2 microns, the size of a small bacterium Light microscopes can magnify effectively to about 1,000 times the size of the actual specimen. At higher magnifications, the image blurs. Fig. 7.1 Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Fig. 7.1 Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Techniques developed in the 20th century have enhanced contrast and enabled particular cell components to be labeled so that they stand out. While a light microscope can resolve individual cells, it cannot resolve much of the internal anatomy, especially the organelles. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings To resolve smaller structures we use an electron microscope (EM), which focuses a beam of electrons through the specimen or onto its surface. Because resolution is inversely related to wavelength used, electron microscopes with shorter wavelengths than visible light have finer resolution. Theoretically, the resolution of a modern EM could reach 0.1 nanometer (nm), but the practical limit is closer to about 2 nm. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Transmission electron microscopes (TEM) are used mainly to study the internal ultrastructure of cells. A TEM aims an electron beam through a thin section of the specimen. The image is focused and magnified by electromagnets. To enhance contrast, the thin sections are stained with atoms of heavy metals. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Scanning electron microscopes (SEM) are useful for studying surface structures. The sample surface is covered with a thin film of gold. The beam excites electrons on the surface. These secondary electrons are collected and focused on a screen. The SEM has great depth of field, resulting in an image that seems three-dimensional. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Electron microscopes reveal organelles, but they can only be used on dead cells and they may introduce some artifacts. Light microscopes do not have as high a resolution, but they can be used to study live cells. Microscopes are a major tool in cytology, the study of cell structures. Cytology coupled with biochemistry, the study of molecules and chemical processes in metabolism, developed modern cell biology. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Isolating Organelles by Cell Fractionation Cell fractionation Takes cells apart and separates the major organelles from one another The centrifuge Is used to fractionate cells into their component parts Isolation of organelles to study their functions The goal of cell fractionation is to separate the major organelles of the cells so that their individual functions can be studied. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings This process is driven by a ultracentrifuge, a machine that can spin at up to 130,000 revolutions per minute and apply forces more than 1 million times gravity (1,000,000 g). Fractionation begins with homogenization, gently disrupting the cell. Then, the homogenate is spun in a centrifuge to separate heavier pieces into the pellet while lighter particles remain in the supernatant. As the process is repeated at higher speeds and longer durations, smaller and smaller organelles can be collected in subsequent pellets. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Cell fractionation prepares quantities of specific cell components. This enables the functions of these organelles to be isolated, especially by the reactions or processes catalyzed by their proteins. For example, one cellular fraction is enriched in enzymes that function in cellular respiration. Electron microscopy reveals that this fraction is rich in the organelles called mitochondria. Cytology and biochemistry complement each other in connecting cellular structure and function. Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings The Cell A cell is the smallest unit of living matter. Don’t confuse this with: atom, element, proton, etc. Cell Theory Who? Matthias Schleiden, Theodor Schwann, Rudolf Virchow When? 1800s What does it say? All organisms are made of cells. A cell is the structural & function unit of organs. All cells come from pre-existing cells. Cells are capable of self-reproduction. Cell Size Types of Cells Unicellular organisms Bacteria, Protists, etc. Multicellular organisms Plants Animals Muscles, skin, nerves, liver, digestive, bones, blood, immune system, lungs, etc. How do we know what happens in each part of the cell? Radioisotopes are used to "trace" different chemical reactions through a cell. Separate cellular structures with a blender Centrifuge material and analyze each layer. Two basic cell types Eukaryotes (Eu = true) (kary = nucleus) Organisms whose cells contain a membrane-bound nucleus and other organelles. Prokaryotes (Pro = before) Organisms without a membrane-bound nucleus (bacteria). * These cells have genetic information, but not in a nucleus. * Evolutionists chose the prefix “pro” because they believe these evolved before others. A Typical Cell 1. 2. 3. 4. Has structural components - cytoskeleton (made up of protein filaments) Has organelles that perform specific functions in the cell (Mitochondria -produces energy) Has certain active genes to give the cell a specific function (heart cells, liver cells, brain cells, etc) Has the information to perpetuate the whole organism as well as its specific function (Nucleus - DNA) Prokaryotic Cells Organisms with prokaryotic cells are called “prokaryotes” Prokaryotes have no true nucleus or organelles. Have a single strand of “looped” DNA Most prokaryotes are single-celled microscopic organisms. Eukaryotic Cells 1. Organisms composed of eukaryotic cells are called “eukaryotes” 2. Have a membrane bound nucleus which contains the cell’s DNA 3. Some eukaryotes are one-celled organisms 4. All multicellular organisms are eukaryotes 5. Have organelles, each of which is surrounded by (or bound in) a “plasma membrane” Some Example Prokaryotes Coccusshaped bacterium Bacillusshaped bacterium Spirillumshaped bacterium Prokaryotes vs. Eukaryotes (1) Size Prokaryotes ≤ 10 µm example: Bacteria & Archea Eukaryotes ≥ 10 µm example: Protista, Fungi, Plants, Complexity Prokaryotes – simple Eukaryotes – complex Location of chromosomes Animals Prokaryotes – free in cytosol Eukaryotes – within a membrane-bound nucleus Flagellar mechanisms differ Prokaryotes vs. Eukaryotes (2) Very simple cells Always single-celled No nucleus DNA arranged in one single loop Found only in kingdom Monera (bacteria) Complex cells Can be singlecelled or multicellular Have a nucleus DNA arranged in many separate strands Found in Animal, Plant, Protists, and Fungi kingdoms Prokaryotic Cells 1. 2. 3. 4. Have no membrane-bound organelles Include true bacteria On earth 3.8 million years Found nearly everywhere 1. Naturally in soil, air, 2. Hot springs nucleoid (DNA) Prokaryotic Cells ribosomes food granule prokaryotic flagellum plasma membrane cytoplasm cell wall Viruses 1. Viruses contain DNA or RNA & a protein coat 2. Some are enclosed by an envelope 3. Most viruses infect only specific types of cells in one host 4. Host range is determined by specific host attachment sites and cellular factors Comparison of Cells and Viruses Bacterium (prokaryote) Animal (eukaryote) Plant (eukaryote) Prokaryotic bacteria cells surrounding a eukaryotic cell (possibly a white blood cell?) Comparison between prokaryotes & eukaryotes (1) Prokaryotes Eukaryotes Typical organisms Bacteria, archaea Protists, fungi, plants, animals Typical size 1 - 10 mm 10 – 100 mm Type of nucleus Nucleoid, no real membrane Real nucleus w/ double membrane DNA Circular (usually) Linear molecules (chromosomes) with histone proteins RNA/protein synthesis Coupled in cytoplasm RNA synthesis inside the nucleus, protein synthesis in cytosol Comparison between prokaryotes & eukaryotes (2) Prokaryotes Eukaryotes Ribosomes 50S + 30S 60S + 40S Cytoplasmic structure Very few structures Highly structured by endomembraes and a cytoskeleton Cell movement Flagella Flagella & cilia made of tubulin Mitochondria None One to several dozen Chloroplasts None Algae & plants Comparison between prokaryotes & eukaryotes (3) Prokaryotes Eukaryotes Organization Usually single cells Single cells, colonies, higher multicultural organisms w/ specialized cells Cell division Binary fission (simple division) Mitosis & meiosis Eukaryotic Cell Cell Structure & Function (1) Bacteria Cell Structure & Function (2) Typical Plant Cell Structure & Function (3) Generic Animal Cell Eukaryotic Cells Structure (1) 1. Have numerous internal structures 2. Various types & forms 3. Plants, animals, fungi, protists 4. Multicellular organisms Eukaryotic Cells Structure (2) 1. The cell consists of two main compartments: 1. The nuclear 2. The cytoplasmic 2. The nucleus contains the genetic information that regulates the structure and function of all eukaryotic cells 3. The cytoplasm contains numerous cellular organelles, which perform specific functions Plant & Animal Cells (1) Similarities 1. Both constructed from eukaryotic cells 2. Both contain similar organelles 3. Both surrounded by cell membrane Plant & Animal Cells (2) Differences Plants have 1. Cell wall – provides strength & rigidity 2. Have chloroplasts, photosynthetic site 3. Large vacuoles Animals have 1. Other organelle not found in plants (lysosomes formed from Golgi) 2. Centrioles, important in cell division Cellular Organelles Cytoplasm Nucleus Chromosomes, nuclear envelope, nuclear pores, nucleolus Ribosomes Endoplasmic reticulum (smooth & rough) Golgi Apparatus Lysosomes Vesicles Peroxisomes Vacuoles Chloroplast Mitochondria Cytoskeleton Centrioles Cilia, Flagella Plasma Membrane Nucleus The nucleus is separated from the cytoplasm by the nuclear envelope Nucleus Structure Nucleus: DNA stored here. Nuclear The Control Center envelope: membrane surrounding the nucleus Nuclear pores: open portals of communication between the nucleus & cytoplasm Chromatin: condensed DNA Chromosome: very tightly packed DNA Nucleolus: dense region of chromatin DNA proteins 1. DNA is associated with two major types of proteins: 2. The histone and nonhistone chromosomal proteins 3. The histones are primarily structure molecules that pack DNA into chromatin fibers 4. The nonhistones include proteins that carry out one of the most important cellular functions, the regulation of gene activity Chromosomes 1. 2. 3. 4. 5. DNA molecule, with its associated histone and nonhistone proteins, is a chromosome There are five classes of histone proteins: H1 H2A H2B H3 H4 Nucleosomes H2A, H2B, H3, and H4 are called core histones because they form a beadlike core structure around which DNA wraps to form nucleosomes. H1 is called the linker Human Chromosomes The entire complement of 46 chromosomes in a human cell, has a total length of about 1 meter Nucleolus (Nucleoli) The RNA of ribosomes is synthesized from genes in the nucleolus No membranes separate nucleoli from the surrounding chromatin in the nucleus Protein-encoding gene Each DNA segment containing the information in a protein constitutes a gene The information in a Protein-encoding gene is copied into a messenger RNA (mRNA) molecules that moves to the cytoplasm through the pores of the nuclear envelop In the cytoplasm, mRNA molecules are used by ribosomes as directions for the assembly of proteins DNA -----------> mRNA -----------> Protein (enzymes) RNA types 1. mRNA 2. rRNA 3. tRNA Ribosomes: protein factories Rough ER: make proteins (studded with ribosomes) Smooth ER: make lipids, modify proteins made in RER Mitochondria & Chloroplast: Power Stations of the cell Mitochondria (1) 1. The mitochondria major role is ATP production in the eukaryotic cell 2. These are mobile and flexible organelles 3. Although in some cells they tend to stay in a fixed position 4. Mitochondria are also self-reproducing, they have their own circular DNA Mitochondria (2) 1. Generate cellular energy in the form of ATP molecules 2. ATP is generated by the systematic breakdown of glucose = cell respiration 3. Also, surrounded by 2 membrane layers 4. Contain their own DNA! 5. A typical liver cell may have 1,700 mitochondria 6. All your mitochondria come from your mother Inner Membrane and matrix (3) Electron transport system Oxidative phosphorylation (4) H+ H+ 3H+ H+ ATP synthase IV III I H+ H+ 3 ATP H+ 2e- NADH 3H+ 3 ADP + 3 Pi Inner Membrane Electron transport system Mitochondria Chloroplasts Compartments 2 3 pH 7–8 5-8 Metabolic Sites Matrix: TCA cycle, ATP synthesis ETC: 3H+ pumps Stroma: Calvin cycle & ATP synthesis ETC: 1H+ pump Substrates Oxidizes glucose, other metabolites to make ATP Light Rxn: use energy from light to synthesize NADPH & ATP Dark Rxn: use CO2 & H2O & NADPH & ATP to synthesize glucose Wastes CO2 & H2O O2 Chloroplast Mitochondria Endoplasmic reticulum (1) 1. Rough endoplasmic reticulum 2. smooth endoplasmic reticulum are connected and are continuous with the nuclear envelope Rough endoplasmic reticulum (2) 1. It is rough because imbedded in the membrane are ribosomes 2. The site of the synthesis of secretory proteins 3. The rough ER is also the site for the synthesis of membrane 4. Enzymes synthesize phospholipid that forms all the membranes of the cell 5. Ribosomes in the rough ER synthesize protein that then are converted to glycoprotein and packaged in transport vesicles for secretion Smooth endoplasmic reticulum (3) 1. The smooth ER is the site for the synthesis of lipids, phospholipids, and steriods 2. Note that the production of steriod hormones is tissue specific 3. For example, it is the smooth ER of the cells of the ovaries and testes that synthesize the sex hormones 4. The smooth ER of the liver has several additional functions Smooth endoplasmic reticulum (4) 5. Enzymes in the smooth ER regulate the release of sugar into the bloodstream 6. Other enzymes break down toxic chemicals 7. As the liver is exposed to additional doses of a drug the liver increases the amount of smooth ER to handle it 8. It then takes more drug to get past the detoxifiying ability of the liver 9. Finally the smooth ER functions to store calcium ions Golgi apparatus 1. The Golgi apparatus, like the ER, is a series of folded membranes 2. It functions in processing enzymes and other products of the ER to a finished product 3. It is the source of the production of lysosomes 4. Receives proteins & lipids in membrane-bound vesicles from ER 5. Modifies those proteins & lipids 6. Sorts and ships the proteins & lipids away in membrane-bound vesicles vesicles from ER vesicles leaving Golgi complex Golgi complex Lysosomes 1. These are membrane bound vesicles that harbor digestive enzymes 2. The membrane of a lysosome will fuse with the membrane of vacuoles releases these digestive enzymes to the interior of the vacuole to digest the material inside the vacuole Vacuoles 1. These are membrane-bound sacs that have many different functions 2. The central vacuole of a plant cell serves as a large lysosome 3. It may also function in absorbing water 4. The central vacuoles of flower petal cells may hold the pigments that give the flower its color Endomembrane system This section reviews the endomembrane system which encludes the nuclear envelope, the rough and smooth ER, the Golgi apparatus, lysosomes and vacuoles Ribosomes 1. Ribosomes assemble amino acid monomers into polypeptide chains 2. Associated with the ER 3. Composed of RNA and proteins rough endoplasmic reticulum ribosomes 0.5 micrometers smooth endoplasmic reticulum 0.5 micrometers vesicles Ribosome Assembly/Structure If individual proteins and rRNAs are mixed, functional ribosomes will assemble Structures of large and small subunits have been determined in 2000/2001 Growing peptide chain is thought to thread through the tunnel during protein synthesis Eukaryotic ribosomes Mitochondrial and chloroplast ribosomes are quite similar to prokaryotic ribosomes, reflecting their supposed prokaryotic origin Cytoplasmic ribosomes are larger and more complex, but many of the structural and functional properties are similar Mechanics of protein synthesis 1. All protein synthesis involves three phases: initiation, elongation, termination 2. Initiation involves binding of mRNA and initiator aminoacyl-tRNA to a small subunit, followed by binding of a large subunit 3. Elongation: synthesis of all peptide bonds with tRNAs bound to acceptor (A) and peptidyl (P) sites 4. Termination occurs when "stop codon" reached Cell Motility The movement of whole cells is made possible through the membrane pliability and the rearrangement of the cytoskeleton and internal components Cytoskeleton provides strength, flexibility and motility Cytoskeleton Eukaryotic cells has a meshwork of tiny fibers that support the structure. This network is the cytoskeleton. Three types of fibers exist. 1. Microfilaments are solid helical rods composed of the protein actin. There is a twist double chain of actin molecules that make up microfilaments. These are found in cells that must contract such as muscle cells. 2. Intermediate filaments are variable but in general are ropelike structures made of twisted filaments of fibrous proteins. These function in bearing tension and anchoring organelles. 3. Microtubles are straight, hollow tubes composed of proteins called tubulins. These anchor organelles and provide tract along which organelles may move. They also make up flagella and cilia. 1. Microfilaments(6 nm) Functions 1. Anchor cytockeleton to integral proteins 2. Determine the consistency of the cytoplasm, 3. Interacts with myosin to produce movement 2. Intermediate filaments (7-11 nm) (Protein composition varies between cell types) Functions 1. Strengthen cell and help maintain shape 2. Stabilize the position of organelles 3. Stabilize the position of the cell with respect to surrounding cells thru specialized membrane attachments 3. Microtubules (up to 25 nm) all cells contain microtubules, made up of protein tubulin. Largest cytoskeletal component Functions 1. Primary cytoskeletal component 2. Disassembly of microtubules provides a mechanism for changing the shape of the cell and assisting in movement 3. Used to transport other proteins around the cell in association with motor proteins kinesin and dynein 4. Forms spindle apparatus during cell division 5. Form structural cell components such as cilia and centrioles The Cytoskeleton Figure 3.5 Cilia and flagella These are found on cells, such as protists, that are motile. Cilia are short and numerous. Longer less numerous appendages are flagella. These are composed of a core of microtubules wrapped in an extension of the plasma membrane. It is sufficient to know that Energy is required to move the cilia or flagella in a whiplike motion to propel the cell. Cell surfaces Cells are held tightly together is higher organisms. There is also a considerable amount of cell communication for lack of a better term. Cell junctions are structures that hold cells together. There are three types. Tight junctions bind cells together forming a leakproof sheet. Anchoring junctions attach adjacent cells or cells to an extracellular matrix (the substance in which tissues cells are embedded. These are leaky compared to tight junctions. Communicating junctions are channels between similar cells. Plasmodesmata are passages between adjacent plant cells that allow material to go from one cell to the next. Communication junctions fulfill the same role between animal cells. Cytoskeleton: provides structure and Support for the cell. Also provides a Scaffolding for vesicle transportation Rapid Review (1) Organelle Prokaryotes Eukaryotes Functions Animal cells Plant cells + _ + Protects & shapes the cell + Plasma membrane + + Selective barrier consisting of bilayers of phospholipids, proteins, & CHO + + + Protein synthesis, formed in nucleolus Cell wall Ribosome Rapid Review (2) Organelle Prokaryotes Eukaryotes Animal cells Plant cells Functions Sooth ER _ + + Lipid synthesis, detoxification, CHO metabolism, no ribosomes on cytoplasmic surface Rough ER _ + + Synthesizes proteins to secrete or send to plasma membrane. Contains ribosomes on cytoplasmic surface Gogli _ + + Modifies lipids, proteins, etc & sends them to other sites in the cell Rapid Review (3) Organelle Prokaryotes Eukaryotes Functions Animal cells Plant cells Mitochondria _ + + Powerhouse of cell; host major energy-producing steps of respiration Lysosome _ + + Contains enzymes that digest organic compounds; serves as cell’s stomach Nucleus _ + + Control center of cell. Host for transcription, replication & DNA Rapid Review (4) Organelle Prokaryotes Eukaryotes Functions Animal Plant cells cells Peroxisome _ + + Breakdown of FA, detoxification of alcohol Chloroplast _ _ + Site of photosynthesis Vacuole _ + (small) + (large) Storage vault of cells Rapid Review (5) Organelle Prokaryotes Eukaryotes Functions Animal Plant cells cells Cytoskeleton _ + + Consists of microtubules (cell division, cilia, flagella), microfilaments (muscles), & intermediate filaments (reinforcing position of organelles _ + _ Part of microtubule separation apparatus that assists cell division Centrioles Plasma Membrane Boundary that separates the living cell from it’s nonliving surroundings. Phospholipid bilayer Amphipathic - having both: hydrophilic heads hydrophobic tails ~8 nm thick Phospholipid Plasma Membrane - cont. Controls traffic into and out of the cell with phospholipids and transport proteins. Selectively permeable Transport protein The Permeability of the Plasma Membrane The property of biological membranes which allows some substances to cross more easily than others. The plasma membrane is differentially permeable. Macromolecules cannot pass through because of size, and tiny charged molecules do not pass through the nonpolar interior of the membrane. Small, uncharged molecules pass through the membrane, following their concentration gradient. How molecules cross the plasma membrane Movement of materials across a membrane may be passive or active. Passive transport does not use chemical energy; diffusion and facilitated transport are both passive. Active transport requires chemical energy and usually a carrier protein. Exocytosis and endocytosis transport macromolecules across plasma membranes using vesicle formation, which requires energy. Fluid Mosaic 1972 - Singer and Nicolson called the membrane a “Fluid Mosaic Model”. Mosaic: Fluid: proteins and phospholipids can move freely in the membrane. different proteins embedded in the phospholipids. Fluid-mosaic model of membrane structure Fluid Mosaic - cont. Components of a phospholipid bilayer. 1. phospholipids 2. proteins - enzymes, receptors, transport. 3. glycolipids 4. glycoproteins 5. carbohydrates 6. cholesterol Cells live in fluid environments, with water inside and outside the cell. Hydrophilic (water-loving) polar heads of the phospholipid molecules lie on the outwardfacing surfaces of the plasma membrane. Hydrophobic (water-fearing) nonpolar tails extend to the interior of the plasma membrane. Plasma membrane proteins may be peripheral proteins or integral proteins. Aside from phospholipid, cholesterol is another lipid in animal plasma membranes; related steroids are found in plants. Cholesterol strengthens the plasma membrane. When phospholipids have carbohydrate chains attached, they are called glycolipids. When proteins have carbohydrate chains attached, they are called glycoproteins. Carbohydrate chains occur only on the exterior surface of the plasma membrane. The outside and inside surfaces of the plasma membrane are not identical. In animal cells, the carbohydrate chains of cell recognition proteins are collectively called the glycocalyx. The glycocalyx can function in cell-to-cell recognition, adhesion between cells, and reception of signal molecules. The diversity of carbohydrate chains is enormous, providing each individual with a unique cellular “fingerprint”. Osmosis The movement of water across selectively permeable membranes. The water moves from a high concentration to low concentration. Osmosis The diffusion of water across a differentially permeable membrane due to concentration differences is called osmosis. Diffusion always occurs from higher to lower concentration. Water enters cells due to osmotic pressure within cells. Osmosis in cells A solution contains a solute (solid) and a solvent (liquid). Cells are normally isotonic to their surroundings, and the solute concentration is the same inside and out of the cell. “Iso” means the same as, and “tonocity” refers to the strength of the solution. Osmosis in plant and animal cells Question: What’s in a Solution? Answer: solute + solvent solution NaCl H2 0 saltwater + Hypertonic A solution with a greater solute concentration compared to another solution. 3% NaCl 97% H2O Red Blood Cell solution 5% NaCl 95% H2O Hypertonic solutions cause cells to lose water. “Hyper” means more than; hypertonic solutions contain more solute. Animal cells undergo crenation (shrivel) in hypertonic solutions. Plant cells undergo plasmolysis, the shrinking of the cytoplasm. Hypotonic A solution with a lower solute concentration compared to another solution. 3% Na 97% H2O Red Blood Cell solution 1% Na 99% H2O Hypotonic solutions cause cells to swell and possibly burst. “Hypo” means less than. Animal cells undergo lysis in hypotonic solution. Increased turgor pressure occurs in plant cells in hypotonic solutions. Plant cells do not burst because they have a cell wall. Isotonic A solution with an equal solute concentration compared to another solution. 3% Na 97% H2O Red Blood Cell solution 3% Na 97% H2O Movement of H2O Water will “ALWAYS” diffuses down a concentration gradient from a HYPOTONIC solution to a HYPERTONIC solution. “ALWAYS REMEMBER” HYPOTONIC HYPERTONIC Animal Cells Animal cells placed into a hypotonic solution will HEMOLYSIS (EXPLODE). Animal cells placed into a hypertonic solution will CRENATE (SHRIVEL). Hemolysis Crenation Red Blood Cells Plant Cells Firmness or tension (vacuole full) that is found in plant cells (cell wall) that are in a hypotonic environment is called TURGID. This process is called TURGOR PRESSURE. Water Cell Wall Water Central Vacuole Water Plant Cells When the plasma membrane pulls away from the cell wall (vacuole empty) in a hypertonic environment (loss of water) is called PLASMOLYSIS. Water Water plasma membrane Cell Wall Water Transport Proteins Transports molecules or ions across biological membranes 3 types of transport proteins: 1. uniport 2. symport 3. antiport Uniport Transport Protein Carries a single solute across the membrane. extracellular fluid intracellular fluid Symport Transport Protein Translocate 2 different solutes simultaneously in same direction. extracellular fluid intracellular fluid Antiport Transport Protein Exchanges 2 solutes by transporting them in opposite directions. extracellular fluid intracellular fluid Diffusion The net movement of a substance (molecules) down a concentration gradient from an area of high concentration to an area of low concentration. passive transport: expended. facilitated diffusion: type of passive transport which uses transport proteins. NO energy is Diffusion Diffusion is the passive movement of molecules from a higher to a lower concentration until equilibrium is reached. Gases move through plasma membranes by diffusion. Transport by Carrier Proteins Some biologically useful molecules pass through the plasma membrane because of channel proteins and carrier proteins that span the membrane. Carrier proteins are specific and combine with only a certain type of molecule. Facilitated transport and active transport both require carrier proteins. Facilitated transport During facilitated transport, substances pass through a carrier protein following their concentration gradients. Facilitated transport does not require energy. The carrier protein for glucose has two conformations and switches back and forth between the two, carrying glucose across the membrane. Facilitated diffusion of glucose Active Transport The movement of molecules (small or large) across the plasma membrane in which energy (ATP) is required. Examples: 1. 2. 3. Sodium (Na) - Potassium (K) Pump Exocytosis Endocytosis Sodium-Potassium Pump The mechanism that uses energy (active transport) released from splitting ATP to transport Sodium (Na+) out of and Potassium (K+) into cells. extracellular fluid intracellular fluid K+ K+ Na+ Na+ Active transport Carrier proteins involved in active transport are called pumps. The sodium-potassium pump is active in all animal cells, and moves sodium ions to the outside of the cell and potassium ions to the inside. The sodium-potassium pump carrier protein exists in two conformations; one that moves sodium to the inside, and the other that moves potassium out of the cell. The sodium-potassium pump Question: How are large molecules transported into and out of the plasma membranes? Answer: Exocytosis and Endocytosis Exocytosis Cell secretes macromolecules (proteins and other biochemicals) out of cell. Part of the Endomembrane System: the fusion of transport vesicles with plasma membrane. Exocytosis and Endocytosis During exocytosis, vesicles fuse with the plasma membrane for secretion. Some cells are specialized to produce and release specific molecules. Examples include release of digestive enzymes from cells of the pancreas, or secretion of the hormone insulin in response to rising blood glucose levels. Exocytosis Endocytosis The energy requiring movement of particles (foreign or natural) into the cell. 3 types of endocytosis: A. Phagocytosis B. Pinocytosis C. Receptor-mediated endocytosis Endocytosis During endocytosis, cells take in substances by invaginating a portion of the plasma membrane, and forming a vesicle around the substance. Endocytosis occurs as: Phagocytosis – large particles Pinocytosis – small particles Receptor-mediated endocytosis – specific particles Receptor-mediated endocytosis A. Phagocytosis Cell eating: cells engulf particles with pseudopodia and pinches off a food vacuole. Two examples: 1. White Blood Cell 2. Amoeba Food Vacuole Bacteria White Blood Cell Phagocytosis B. Pinocytosis Cell drinking: droplets of extracellular fluid are absorbed into the cell by small vesicles. Example: 1. Fungi Hyphae Food Particles Pinocytosis C. Receptor-Mediated Endocytosis Importing specific macromolecules (hormones) into the cell by the inward budding of vesicles formed from coated pits (receptors). Liver Cell Hormones Receptors Types eucaryotic cells 1. 2. 3. 4. 5. Stem Cells Hemopoietic cells Monocytes Macrophages Phagocytes What are stem cells ? Stem = Root = Source Stem cells are the source of all cells Some stem cells can become any cell type Eventually differentiate to 200 cell types Very complex, poorly understood process Several stages of differentiation Stem, blood, red blood, . . . Reversible? Yes, no, maybe, perhaps, don’t know Where are the Stem Cells ? Two choices – Embryo & Adult Embryo 1. Start with one single cell - the fertilized egg 2. Most flexible - can become any cell type 3. Two choices of embryo source Fertilized in the usual way (Man & Woman) Cloned (use your own cells) Adult 1. Present in normal tissues, including brain 2. Have significant and poorly understood limitations Getting Embryonic Stem Cells Need an embryo Old fashioned way - With egg & sperm In Vitro Fertilization Most common Excess embryos (usually dozens created) Never implanted, so not aborted Fetal (Abortion) – Used for some R&D Therapeutic Cloning More exciting – allows use of own cells Needed to eliminate rejection Not reproductive cloning Move in Congress to ban therapeutic cloning Getting Embryonic Stem Cells Egg divides - first 2 cells, then 4, then 8 . . Becomes “Blastocyst” Hollow ball with key cells inside Harvest at 3-5 days – 30 cells in inner mass Embryo is destroyed in process, key ethical issue Two crucial embryonic cell characteristics Not yet differentiated (pluripotent) Can divide forever (key to culturing) Adult Stem Cells 1. Extracted in VERY small amounts from normal tissues 2. Key to avoiding rejection issues – your own cells 1. 2. Traditional rejection Graft vs. Host Disease – The Stems Attack! 3. Key to avoiding embryonic ethical issues 4. Always partially differentiated (for now) 1. BUT, research points to plasticity 1. Liver stem cells to brain, muscle, and liver 2. Bone marrow to muscle 5. 6. 7. 8. Brain stem cells to blood and muscle Not found in many key organs (Pancreas) Difficult to grow in culture; can’t mass produce Holy Grail-Is there a real master adult stem cell Using Stem Cells 1. Reverse damage 1. Central Nervous System (CNS) damage 1. 2. 3. 4. Parkinson’s Disease Multiple Sclerosis Heart Damage, Diabetes, Bone/Joint loss Cancer - Bone marrow transplant avoidance 2. Fix genetic problems 1. Severe Combined Immune Deficiency (SCID) 2. Chronic Granulomatous Disease (CGD) 3. Make good cells as needed 4. Make whole organs as needed 1. Kidney, Liver, Pancreas, maybe a Thumb Recent Research 1. 2. Three ongoing key focal points Understand basic processes, especially differentiation 1. 2. 3. 4. 5. 6. 7. 8. 3. Differentiation triggers Expand uses (usually embryonic & cloning) Brain – Lots of progress, especially in dopamine, neurons Blood – Lots of immune system work Bones / Joints Pancreas (Diabetes) Liver – Cells come from marrow? Weekly announcements – eyes, veins, heart muscle, skin Improve Adult Cells - Transdifferentiating / Plasticity 1. 2. 3. Found a receptor/protein that limits plasticity (GCNF) Stanford found in Sept they cannot make other cells from adult blood stem cells How do adult cells work – fusion, division; Cancer risk? Treatment Progress 1. 2. Modest to date – Very early in clinical process Parkinson’s injections 1. 2. 3. Multiple Sclerosis 1. 4. Treat by killing own rogue stem cells Israel in June 1. 2. 5. Mixed, but promising results Last week - Got neural stem cells to make dopamine 22 month old infant with immune system failure (SCID) Cured via repaired bone marry stem cells Mouse research this year 1. 2. 3. Extract and fix cell DNA Clone cells to get embryonic stem cells Inject new stem cells to fix immune system Fixing an Immune System Baby with damaged immune system 1. 2. 3. 4. 5. 6. Remove bone marrow stem cells Fix genetic problem in DNA Culture cells to get useful volume Inject new immune cells into baby Normal immune system results !! Works when you have stem cells Mouse Immune Research - Cloned embryonic cells 1. 2. 3. 4. 5. 6. 7. Remove ANY cell Fix DNA problem Clone cells to produce embryo/blastocyst Harvest stem cells Coax to differentiate into immune cells Inject new cells Crucial when body has no usable stem cells Ethics – What’s Wrong ? Most issues surround Embryonic Stem Cells Embryo cannot survive cell extraction When does life begin? What is an Embryo? Does it matter if it’s cloned (no sperm)? Does it matter if not implanted (not aborted)? Is it okay if IVF embryos discarded anyway? Abortions also a source and worry Will we harvest humans? Already having kids for marrow matching But, must abort for stem cells (cord blood?) Future of Stem Cells Immense promise, though much work remains Therapies beginning to emerge – SCID, Parkinson’s Embryonic cells from cloning Adult stem cells will work eventually Eliminate need for embryonic cells & ethical issues Tremendous progress with applications No actual embryo, reduced ethical issues Repaired genetic defects, maybe in womb New therapies for non-genetic diseases (Heart) New organs Repairing genetic defects will be common Change brown hair to blonde ? Ethics, ethics, ethics . . . Hemopoietic cells 1. The basis of hemopoiesis is a small population of self-replicating stem cells, which ultimately can generate all types of blood cells. 2. The process of hematopoiesis is controlled by a group of at least 11 growth factors. 3. Three of these glycoproteins initiate the differentiation of macrophages from uni- and bipotential progenitor cells in the bone marrow. Macrophages and monocytes Their development takes in the bone marrow and passes through the following steps: stem cell committed stem cell monoblast promonocyte monocyte (bone marrow) monocyte (peripheral blood) macrophage (tissues) Blood monocytes The blood monocytes are young cells that already possess migratory, chemotactic, pinocytic and phagocytic activities, as well as receptors for IgG Macrophages Macrophages can be divided into normal macrophages and inflammatory macrophages. Normal macrophages includes macrophages in connective tissue . Inflammatory macrophages are present in various exudates. Phagocytes and since they are derived exclusively from monocytes they share similar properties. Phagocytes Phagocytes are cells which ingest particles. The process of eating particles is called "phagocytosis," a process which is one of the distinguishing features of eukaryotic cells, The Cell Cycle Cell division increases the number of somatic (body) cells, and consists of: Mitosis (division of nucleus) Cytokinesis (division of cytoplasm) Apoptosis (cell death) decreases the number of cells. Both cell division and apoptosis occur during normal development and growth. The cell cycle is an orderly sequence of events that occurs from the time when a cell is first formed until it divides into two new cells. Most of the cell cycle is spent in interphase. Following interphase, the mitotic stage of cell division occurs. The stages of interphase G1 stage – cell growth, cell doubles its organelles, accumulates materials for DNA synthesis. S stage – DNA synthesis occurs, and DNA replication results in duplicated chromosomes. G2 stage – cell synthesizes proteins needed for cell division The cell cycle The Mitotic Stage Following interphase is the M stage, including mitosis and cytokinesis. During mitosis, sister chromatids of each chromosome separate, and become the nuclei of the two daughter cells. The cell cycle ends when cytokinesis, the cleaving of the cytoplasm, is complete. The cell cycle is controlled at three checkpoints: 1. During G1 prior to the S stage 2. During G2 prior to the M stage 3. During the M stage prior to the end of mitosis DNA damage can also stop the cell cycle at the G1 checkpoint. Apoptosis Apoptosis is programmed cell death. Apoptosis occurs because of two sets of enzymes called capsases. The first set, the “initiators” receive a signal to activate the second set, the “executioners”. The second set of capsases activate enzymes that tear apart the cell and its DNA. Maintaining the Chromosome Number When a eukaryotic cell is not dividing, the DNA and associated proteins is a tangled mass of thin threads called chromatin. At the time of cell division, the chromatin condenses to form highly compacted structures called chromosomes. Each species has a characteristic number of chromosomes. Overview of Mitosis The diploid number of chromosomes is found in the somatic (non-sex) cells. The diploid (2n) number of chromosomes contains two chromosomes of each kind. The haploid (n) number of chromosomes contains one chromosome of each kind. In the life cycle of many animals, only sperm and eggs have the haploid number of chromosomes. The nuclei of somatic cells undergo mitosis, a nuclear division in which the number of chromosomes stays constant. Before nuclear division occurs, DNA replication takes place, duplicating the chromosomes. A duplicated chromosome is made of two sister chromatids held together in a region called the centromere. Sister chromatids are genetically identical. At the end of mitosis, each chromosome consists of a single chromatid. During mitosis, the centromeres divide and then the sister chromatids separate, becoming daughter chromosomes. Following mitosis, a 2n parental cell gives rise to two 2n daughter cells, or 2n → 2n. Mitosis occurs when tissues grow or when repair occurs. Following fertilization, the zygote divides mitotically, and mitosis continues throughout the lifespan of the organism. Chromosomes (n) n refers to the number of pairs (or the number of different types of chromosomes) 2n refers to the total number of chromosomes In humans n = 23 In chimpanzees n = 24 In the king crab n = 104 More on Chromosomes Humans reproduce sexually Sexual reproduction means simply that the offspring receive genetic material from BOTH the mother and father Not all organisms reproduce this way For many organisms the offspring receive genetic material from only 1 individual Meiosis 1. Meiosis is the process where the genetic material is reduced to half (i.e. only ONE set of chromosomes per daughter cell) 2. Because Meiosis creates cells with only half the “normal” number of chromosomes, (gametes), when these cells join (as in fertilization) the new individual has the same number of chromosomes as each parent. Meiosis 3. So in fact not ALL human cells have the 23 pairs of chromosomes 4. Humans gametes have 1 copy of each chromosome, not 23 pairs. 5. Gametes are human reproductive cells 1. Male gametes are sperm 2. Female gametes are eggs Meiosis versus Mitosis Mitosis is essential for cell growth 1. It is the process that distributes the genetic evenly material between two daughter cells, so that each daughter cell is genetically identical to the parent cell. 2. It’s simply a way to make more of the same cell Mitosis Phases of Mitosis 1. 2. 3. 4. 5. Interphase Prophase Metaphase Anaphase Telophase REPEAT (This process produces two identical cells. The process then repeats for each of the new cells.) Mitosis 1. Interphase DNA replication Centrosomes form 3. Metaphase 2. Prophase Chromosomes condense become visible as sister chromatids (Condensation) Centrosomes move away from each other Mitotic spindle forms Centrosomes at opposite poles Chromosomes line up at metaphase plate (centromeres line up) Each chromatid is attached to one of the poles via microtubules 4. Anaphase Each of the sister chromatids moves toward the opposite pole Mitosis 5. Telophase Formation of cleavage furrow Cell divides 2 identical copies New nuclear envelope forms New Interphase begins And the process starts all over Late Interphase Early Prophase Late Prophase Metaphase Anaphase Telophase Homologues versus chromatids Homologous chromosomes One from Mom and One from Dad Have different versions of the same genes Sister chromatids Bound at center by centromere Chromatids are identical Product of DNA replication Phases of Meiosis Interphase Meiosis I Cell spends 90 % of it’s life in this phase (like mitosis) Chromosome replication takes place sister chromatids Prophase I Homologous chromosomes come together (synapsis) Form tetrads This is when Crossing Over occurs (at chiasmata, between NON-SISTER chromatids) Centrosomes move away from each other Meiosis I Metaphase I Chromosomes aligned on Metaphase plate (grouped in homologous pairs) Each chromosome is attached to opposite pole Anaphase I Sister chromatids remain attached at centromeres Chromosomes move to opp. poles Telophase I Chromosomes reach poles Cell starts to cleave Produces 2 haploid cells Each cell is NOT identical (crossing over) Cytokinesis Meiosis I Chromosomes and Chromatids During Meiosis I Begin Interphase After Interphase After Prophase I After Metaphase I After Anaphase I After Telophase I # of Chromosomes 4 4 4 4 4 2 # of Chromatids 4 8 8 8 8 4 Meiosis II Prophase II Spindle forms Chromosomes move toward metaphase plate Metaphase II Chromosomes line up along metaphase plate Anaphase II Sister chromatids move toward opposite poles Telophase II Nuclei form at opposite poles Cytokinesis (division of cell) Produces 4 haploid cells Chromosomes and Chromatids During Meiosis II After Prophase II After Metaphase II After Anaphase II After Telophase II # of Chromosomes 2 2 4 2 # of Chromatids 4 4 4 2 Meiosis II Haploidy and Diploidy A cell that has 1 copy of each chromosome is haploid Human gametes are haploid (n) A cell that has 2 copies of each chromosome are diploid Human somatic cells (body cells) are diploid (2n) The 4 Main differences between Meiosis and Mitosis 1. In Meiosis (prophase I) homologous chromosomes pair up and crossing over occurs (Neither of these happens in Mitosis) 2. In Meiosis, homologous pairs align on opposite sides of the metaphase plate Meiosis (Metaphase I) Mitosis (Metaphase) 3. In Meiosis (Anaphase I) sister chromatids do not separate Meiosis (Anaphse I) Mitosis (Anaphase) 4. Meiosis I separates homologous pairs of chromosomes NOT sister chromatids Meiosis (Anaphase I) Mitosis (Anaphase) Cytokinesis Cytokinesis, or cytoplasmic cleavage, accompanies mitosis. Cleavage of the cytoplasm begins in anaphase, but is not completed until just before the next interphase. Newly-formed cells receive a share of cytoplasmic organelles duplicated during the previous interphase. A cleavage furrow begins at the end of anaphase. A band of actin and myosin filaments, called the contractile ring, slowly forms a constriction between the two daughter cells. A narrow bridge between the two cells is apparent during telophase, then the contractile ring completes the division. Cytokinesis Cell Division in Prokaryotes The process of asexual reproduction in prokaryotes is called binary fission. The two daughter cells are identical to the original parent cell, each with a single chromosome. Following DNA replication, the two resulting chromosomes separate as the cell elongates. Meiosis in humans Humans have 23 pairs of homologous chromosomes, or 46 chromosomes total. Prior to meiosis I, DNA replication occurs. During meiosis, there will be two nuclear divisions, and the result will be four haploid nuclei. No replication of DNA occurs between meiosis I and meiosis II. Meiosis I separates homologous pairs of chromosomes Daughter cells are haploid, but chromosomes are still in duplicated condition. Synapsis occurs during meiosis I Meiosis II separates sister chromatids. The completely haploid daughter cells mature into gametes. Fertilization restores the diploid number of chromosomes during sexual reproduction. Meiosis in Detail Meiosis involves the same four phases seen in mitosis prophase metaphase anaphase telophase The occur during both meiosis I and meiosis II. The period of time between meiosis I and meiosis II is called interkinesis. No replication of DNA occurs during interkinesis because the DNA is already duplicated. Comparison of Meiosis with Mitosis 1.Before mitosis and meiosis, DNA replication occurs only once during the interphase prior to cell division. 2.Mitosis requires one division 3.Meiosis requires two divisions. 4.Two diploid daughter cells result from mitosis 5.Four haploid daughter cells result from meiosis. Comparison of Meiosis with Mitosis (cont) 6.Daughter cells from mitosis are genetically identical to parental cells 7.Daughter cells from meiosis are not genetically identical to parental cells. 8.Mitosis occurs in all somatic cells for growth and repair. 9.Meiosis occurs only in the reproductive organs for the production of gametes. Comparison of Meiosis I to Mitosis Meiosis I: Prophase I - pairing of homologous chromosomes Metaphase I – homologous pairs line up at metaphase plate Anaphase I – homologous chromosomes separate Telophase I – daughter cells are haploid Mitosis: Prophase has no such pairing Metaphase – chromosomes align at metaphase plate Anaphase – sister chromatids separate Telophase – diploid cells Comparison of Meiosis II to Mitosis 1.The events of meiosis II are like those of mitosis except in meiosis II, the nuclei contain the haploid number of chromosomes. 2.At the end of telophase II of meiosis II, there are four haploid daughter cells that are not genetically identical. 3.At the end of mitosis, there are two diploid daughter cells that are identical. The human life cycle requires both mitosis and meiosis In males, meiosis occurs as spermatogenesis and produces sperm. In females, meiosis occurs as oogenesis and produces egg cells. Mitosis is involved in the growth of a child and repair of tissues during life. Spermatogenesis in human males produces four viable haploid sperm. Diploid primary spermatocytes undergo meiosis I to produce haploid secondary spermatocytes. Secondary spermatocytes divide by meiosis II to produce haploid spermatids. Spermatids mature into sperm with 23 chromosomes. During oogenesis, a diploid primary oocyte undergoes meiosis I to produce one haploid secondary oocyte and one haploid polar body. The secondary oocyte begins meiosis II but stops at metaphase II and is released at this stage from the ovary. Meiosis II will be completed only if sperm are present. Following meiosis II, there is one haploid egg cell with 23 chromosomes and up to three polar bodies. Polar bodies serve as a dumping ground for extra chromosomes. Oogenesis In humans, both sperm cells and the egg cell have 23 chromosomes each. Following fertilization of the egg cell by a single sperm, the zygote has 46 chromosomes, the diploid number found in human somatic cells. The 46 chromosomes represent 23 pairs of homologous chromosomes. Summary Cell division increases the number of body cells; Apoptosis decreases cell number. Cells goes through a cell cycle. Each species has a characteristic number of chromosomes. Mitosis 1. produces daughter cells that are identical to the parental cell. 2. has four phases designed to maintain the chromosome number. 3. is used for growth and repair of tissues. Summary continued Meiosis 1. reduces the chromosome number. 2. includes two nuclear divisions. 3. results in non-identical haploid gametes. The human life cycle includes both mitosis and meiosis. The process of meiosis and fertilization in humans and other sexually reproducing organisms result in offspring with new genetic combination. A comparison of mitosis and meiosis A comparison of mitosis and meiosis: summary SEXUAL(MEIOSIS) Figure 14.32. Comparison of meiosis and mitosis. Both meiosis and mitosis initiate after DNA replication, so each chromosome consists of two sister chromatids. In meiosis I, homologous chromosomes pair and then segregate to different cells. Sister chromatids then separate during meiosis II, which resembles a normal mitosis. Meiosis thus gives rise to four haploid daughter cells. Fuente: Cooper, 2000 ATTACHMENT Click after each step to view process PENETRATION UNCOATING HOST FUNCTIONS Transcription Translation VIRAL LIFE CYCLE REPLICATION ASSEMBLY (MATURATION) RELEASE MULTIPLICATION